Western blot analysis of extracts of HeLa cells, using Phospho-HSPB1-S15 antibody (AP0040) at 1:1,000 dilution. HeLa cells were treated by EGF (100ng/ml) for 30 minutes after serum-starvation overnight. Secondary antibody: Goat Anti-Rabbit IgG (H+L) (HRP) (AS014) at 1:10,000 dilution. Lysates/proteins: 25µg per lane. Blocking buffer: 3% BSA. Detection: ECL Basic Kit (RM00020). Exposure time: 5min.
Immunoprecipitation analysis of 200µg extracts of HeLa cells, using 3µg Phospho-HSPB1-S15 pAb (AP0040). Western blot was performed from the immunoprecipitate using Phospho-HSPB1-S15 pAb (AP0040) at a dilition of 1:1000. HeLa cells were treated by EGF (100 ng/ml) at 37°C for 30 minutes after serum-starvation overnight.
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